PD1/PDCD1 Antibody

Western blot analysis of PD1/PDCD1 using anti-PD1/PDCD1 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: human Caco-2 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human COLO-320 whole cell lysates, Lane 4: human K562 whole cell lysates, Lane 5: mouse thymus tissue lysates, Lane 6: mouse RAW246.7 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PD1/PDCD1 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for PD1/PDCD1 at approximately 65 KD. The expected band size for PD1/PDCD1 is at 32 KD.